The obligate intracellular pathogen, Anaplasma phagocytophilum, is the causative agent of human, equine, and canine granulocytic anaplasmosis and tick-borne fever (TBF) in ruminants. A. phagocytophilum has become an emerging tick-borne pathogen in the United States, Europe, Africa, and Asia, with increasing numbers of infected people and animals every year. It has been recognized that intracellular pathogens manipulate host cell metabolic pathways to increase infection and transmission in both vertebrate and invertebrate hosts. However, our current knowledge on how A. phagocytophilum affect these processes in the tick vector, Ixodes scapularis is limited. In this study, a genome-wide search for components of major carbohydrate metabolic pathways was performed in I. scapularis ticks for which the genome was recently published. The enzymes involved in the seven major carbohydrate metabolic pathways glycolysis, gluconeogenesis, pentose phosphate, tricarboxylic acid cycle (TCA), glyceroneogenesis, and mitochondrial oxidative phosphorylation and β-oxidation were identified. Then, the available transcriptomics and proteomics data was used to characterize the mRNA and protein levels of I. scapularis major carbohydrate metabolic pathway components in response to A. phagocytophilum infection of tick tissues and cultured cells. The results showed that major carbohydrate metabolic pathways are conserved in ticks. A. phagocytophilum infection inhibits gluconeogenesis and mitochondrial metabolism, but increases the expression of glycolytic genes. A model was proposed to explain how A. phagocytophilum could simultaneously control tick cell glucose metabolism and cytoskeleton organization, which may be achieved in part by up-regulating and stabilizing hypoxia inducible factor 1 alpha in a hypoxia-independent manner. The present work provides a more comprehensive view of the major carbohydrate metabolic pathways involved in the response to A. phagocytophilum infection in ticks, and provides the basis for further studies to develop novel strategies for the control of granulocytic anaplasmosis.

Institute of Parasitology Biology Center Czech Academy of SciencesCeské Budejovice Czechia; Department of Virology Veterinary Research InstituteBrno Czechia

Institute of Parasitology Biology Center Czech Academy of SciencesCeské Budejovice Czechia; Faculty of Science University of South BohemiaCeské Budejovice Czechia

SaBio Instituto de Investigación en Recursos Cinegéticos Ciudad Real Spain

SaBio Instituto de Investigación en Recursos Cinegéticos Ciudad Real Spain; Department of Veterinary Pathobiology Center for Veterinary Health Sciences Oklahoma State UniversityStillwater OK USA

Zobrazit více v PubMed

Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. (1990). Basic local alignment search tool. J. Mol. Biol. 215, 403–410. 10.1016/S0022-2836(05)80360-2 PubMed DOI

Asanovich K. M., Bakken J. S., Madigan J. E., Aguero-Rosenfeld M., Wormser G. P., Dumler J. S. (1997). Antigenic diversity of granulocytic Ehrlichia isolates from humans in Wisconsin and New York and a horse in California. J. Infect. Dis. 176, 1029–1034. 10.1086/516529 PubMed DOI

Atilgan A. R., Durell S. R., Jernigan R. L., Demirel M. C., Keskin O., Bahar I. (2001). Anisotropy of fluctuation dynamics of proteins with an elastic network model. Biophys. J. 80, 505–515. 10.1016/S0006-3495(01)76033-X PubMed DOI PMC

Augustin R. (2010). The protein family of glucose transport facilitators: it's not only about glucose after all. IUBMB Life 62, 315–333. 10.1002/iub.315 PubMed DOI

Ayllón N., Villar M., Busby A. T., Kocan K. M., Blouin E. F., Bonzón-Kulichenko E., et al. . (2013). Anaplasma phagocytophilum inhibits apoptosis and promotes cytoskeleton rearrangement for infection of tick cells. Infect. Immun. 81, 2415–2425. 10.1128/IAI.00194-13 PubMed DOI PMC

Ayllón N., Villar M., Galindo R. C., Kocan K. M., Šíma R., López J. A., et al. . (2015). Systems biology of tissue-specific response to Anaplasma phagocytophilum reveals differentiated apoptosis in the tick vector Ixodes scapularis. PLoS Genet. 11:e1005120. 10.1371/journal.pgen.1005120 PubMed DOI PMC

Badeaux A. I., Shi Y. (2013). Emerging roles for chromatin as a signal integration and storage platform. Nat. Rev. Mol. Cell. Biol. 14, 211–224. 10.1038/nrm3545 PubMed DOI

Bensaad K., Tsuruta A., Selak M. A., Vidal M. N., Nakano K., Bartrons R., et al. . (2006). TIGAR, a p53-inducible regulator of glycolysis and apoptosis. Cell 126, 107–120. 10.1016/j.cell.2006.05.036 PubMed DOI

Berg J. M., Tymoczko J. L., Stryer L. (2002). Biochemistry, 5th Edn. New York, NY: W H Freeman Press.

Biasini M., Bienert S., Waterhouse A., Arnold K., Studer G., Schmidt T., et al. . (2014). SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information. Nucleic. Acids. Res. 42, W252–W258. 10.1093/nar/gku340 PubMed DOI PMC

Boldrick J. C., Alizadeh A. A., Diehn M., Dudoit S., Liu C. L., Belcher C. E., et al. . (2002). Stereotyped and specific gene expression programs in human innate immune responses to bacteria. Proc. Natl. Acad. Sci. U.S.A. 99, 972–977. 10.1073/pnas.231625398 PubMed DOI PMC

Borrelli K. W., Vitalis A., Alcantara R., Guallar V. (2005). PELE: protein energy landscape exploration. A novel Monte Carlo based technique. J. Chem. Theor. Comput. 1, 1304–1311. 10.1021/ct0501811 PubMed DOI

Burgess S. C., He T., Yan Z., Lindner J., Sherry A. D., Malloy C. R., et al. . (2007). Cytosolic phosphoenolpyruvate carboxykinase does not solely control the rate of hepatic gluconeogenesis in the intact mouse liver. Cell Metab. 5, 313–320. 10.1016/j.cmet.2007.03.004 PubMed DOI PMC

Cabezas-Cruz A., Alberdi P., Ayllón N., Valdés J. J., Pierce R., Villar M., et al. . (2016). Anaplasma phagocytophilum increases the levels of histone modifying enzymes to inhibit cell apoptosis and facilitate pathogen infection in the tick vector Ixodes scapularis. Epigenetics 11, 303–319. 10.1080/15592294.2016.1163460 PubMed DOI PMC

Choi E. Y., Kim E. C., Oh H. M., Kim S., Lee H. J., Cho E. Y., et al. . (2004). Iron chelator triggers inflammatory signals in human intestinal epithelial cells: involvement of p38 and extracellular signal-regulated kinase signaling pathways. J. Immunol. 172, 7069–7077. 10.4049/jimmunol.172.11.7069 PubMed DOI

Dalei W., Nalini P., Jingping L., Youngchang K., Fraydoon R. (2015). Structural integration in hypoxia-inducible factors. Nature 524, 303–308. 10.1038/nature14883 PubMed DOI

de la Fuente J., Estrada-Peña A., Cabezas-Cruz A., Kocan K. M. (2016a). Anaplasma phagocytophilum uses common strategies for infection of ticks and vertebrate hosts. Trends Microbiol. 24, 173–180. 10.1016/j.tim.2015.12.001 PubMed DOI

de la Fuente J., Estrada-Pena A., Venzal J. M., Kocan K. M., Sonenshine D. E. (2008). Overview: ticks as vectors of pathogens that cause disease in humans and animals. Front. Biosci. 13, 6938–6946. 10.2741/3200 PubMed DOI

de la Fuente J., Villar M., Cabezas-Cruz A., Estrada-Peña A., Ayllón N., Alberdi P. (2016c). Tick-host-pathogen interactions: conflict and cooperation. PLoS Pathog. 12:e1005488. 10.1371/journal.ppat.1005488 PubMed DOI PMC

de la Fuente J., Waterhouse R. M., Sonenshine D. E., Roe R. M., Ribeiro J. M., Sattelle D. B. (2016b). Tick genome assembled: new opportunities for research on tick-host-pathogen interactions. Front. Cell. Infect. Microbiol. 6:103. 10.3389/fcimb.2016.00103 PubMed DOI PMC

Déry M. A., Michaud M. D., Richard D. E. (2005). Hypoxia-inducible factor 1: regulation by hypoxic and non-hypoxic activators. Int. J. Biochem. Cell. Biol. 37, 535–540. 10.1016/j.biocel.2004.08.012 PubMed DOI

Dunning Hotopp J. C., Lin M., Madupu R., Crabtree J., Angiuoli S. V., Eisen J. A., et al. . (2006). Comparative genomics of emerging human ehrlichiosis agents. PLoS Genet. 2:e21. 10.1371/journal.pgen.0020021 PubMed DOI PMC

Eisenreich W., Heesemann J., Rudel T., Goebel W. (2013). Metabolic host responses to infection by intracellular bacterial pathogens. Front. Cell. Infect. Microbiol. 3:24. 10.3389/fcimb.2013.00024 PubMed DOI PMC

Eisenreich W., Heesemann J., Rudel T., Goebel W. (2015). Metabolic adaptations of intracellullar bacterial pathogens and their mammalian host cells during infection (“Pathometabolism”). Microbiol. Spectr. 3, 27–58. 10.1128/microbiolspec.MBP-0002-2014 PubMed DOI

Finn R. D., Bateman A., Clements J., Coggill P., Eberhardt R. Y., Eddy S. R., et al. . (2014). Pfam: the protein families database. Nucleic Acids Res. 42, D222–D230. 10.1093/nar/gkt1223 PubMed DOI PMC

Gulia-Nuss M., Nuss A. B., Meyer J. M., Sonenshine D. E., Roe R. M., Waterhouse R. M., et al. . (2016). Genomic insights into the Ixodes scapularis tick vector of Lyme disease. Nat. Commun. 7:10507. 10.1038/ncomms10507 PubMed DOI PMC

Hu C. J., Iyer S., Sataur A., Covello K. L., Chodosh L. A., Simon M. C. (2006). Differential regulation of the transcriptional activities of hypoxia-inducible factor 1 α (HIF-1α) and HIF-2α in stem cells. Mol. Cell. Biol. 26, 3514–3526. 10.1128/MCB.26.9.3514-3526.2006 PubMed DOI PMC

Huang H., Wang X., Kikuchi T., Kumagai Y., Rikihisa Y. (2007). Porin activity of Anaplasma phagocytophilum outer membrane fraction and purified P44. J. Bacteriol. 189, 1998–2006. 10.1128/JB.01548-06 PubMed DOI PMC

Kocan K. M., de la Fuente J., Cabezas-Cruz A. (2015). The genus Anaplasma: new challenges after reclassification. Rev. Sci. Technol. 34, 577–586. 10.20506/rst.34.2.2381 PubMed DOI

Lecuit M., Sonnenburg J. L., Cossart P., Gordon J. I. (2007). Functional genomic studies of the intestinal response to a foodborne enteropathogen in a humanized gnotobiotic mouse model. J. Biol. Chem. 282, 15065–15072. 10.1074/jbc.M610926200 PubMed DOI

Li X. B., Gu J. D., Zhou Q. H. (2015). Review of aerobic glycolysis and its key enzymes–new targets for lung cancer therapy. Thorac. Cancer 6, 17–24. 10.1111/1759-7714.12148 PubMed DOI PMC

Li X., Jacobson M. P., Zhu K., Zhao S., Friesner R. A. (2007). Assignment of polar states for protein amino acid residues using an interaction cluster decomposition algorithm and its application to high resolution protein structure modeling. Proteins 66, 824–837. 10.1002/prot.21125 PubMed DOI

Madden T. L., Tatusov R. L., Zhang J. (1996). Applications of network BLAST server. Methods Enzymol. 266, 131–141. 10.1016/S0076-6879(96)66011-X PubMed DOI

Misra R. M., Bajaj M. S., Kale V. P. (2012). Vasculogenic mimicry of HT1080 tumour cells in vivo: critical role of HIF-1α-neuropilin-1 axis. PLoS ONE 7:e50153. 10.1371/journal.pone.0050153 PubMed DOI PMC

Munderloh U. G., Jauron S. D., Fingerle V., Leitritz L., Hayes S. F., Hautman J. M., et al. . (1999). Invasion and intracellular development of the human granulocytic ehrlichiosis agent in tick cell culture. J. Clin. Microbiol. 37, 2518–2524. PubMed PMC

Pagé E. L., Robitaille G. A., Pouysségur J., Richard D. E. (2002). Induction of hypoxia-inducible factor-1alpha by transcriptional and translational mechanisms. J. Biol. Chem. 277, 48403–48409. 10.1074/jbc.M209114200 PubMed DOI

Pilot-Storck F., Chopin E., Rual J. F., Baudot A., Dobrokhotov P., Robinson-Rechavi M., et al. . (2010). Interactome mapping of the phosphatidylinositol 3-kinase-mammalian target of rapamycin pathway identifies deformed epidermal autoregulatory factor-1 as a new glycogen synthase kinase-3 interactor. Mol. Cell. Proteomics 9, 1578–1593. 10.1074/mcp.M900568-MCP200 PubMed DOI PMC

Shin J. H., Yang J. Y., Jeon B. Y., Yoon Y. J., Cho S. N., Kang Y. H., et al. . (2011). (1)H NMR-based metabolomic profiling in mice infected with Mycobacterium tuberculosis. J. Proteome Res. 10, 2238–2247. 10.1021/pr101054m PubMed DOI

Somashekar B. S., Amin A. G., Rithner C. D., Troudt J., Basaraba R., Izzo A., et al. . (2011). Metabolic profiling of lung granuloma in Mycobacterium tuberculosis infected guinea pigs: ex vivo 1H magic angle spinning NMR studies. J. Proteome Res. 10, 4186–4195. 10.1021/pr2003352 PubMed DOI

Stuen S., Granquist E. G., Silaghi C. (2013). Anaplasma phagocytophilum—a widespread multi-host pathogen with highly adaptive strategies. Front. Cell. Infect. Microbiol. 3:31. 10.3389/fcimb.2013.00031 PubMed DOI PMC

Sultana H., Neelakanta G., Kantor F. S., Malawista S. E., Fish D., Montgomery R. R., et al. . (2010). Anaplasma phagocytophilum induces actin phosphorylation to selectively regulate gene transcription in Ixodes scapularis ticks. J. Exp. Med. 207, 1727–1743. 10.1084/jem.20100276 PubMed DOI PMC

Villar M., Ayllón N., Alberdi P., Moreno A., Moreno M., Tobes R., et al. . (2015). Integrated metabolomics, transcriptomics and proteomics identifies metabolic pathways affected by Anaplasma phagocytophilum infection in tick cells. Mol. Cell. Proteomics 14, 3154–3172. 10.1074/mcp.M115.051938 PubMed DOI PMC

Wang Q., Liang B., Shirwany N. A., Zou M. H. (2011). 2-Deoxy-D-glucose treatment of endothelial cells induces autophagy by reactive oxygen species-mediated activation of the AMP-activated protein kinase. PLoS ONE 6:e17234. 10.1371/journal.pone.0017234 PubMed DOI PMC

Weidemann A., Breyer J., Rehm M., Eckardt K. U., Daniel C., Cicha I., et al. . (2013). HIF-1α activation results in actin cytoskeleton reorganization and modulation of Rac-1 signaling in endothelial cells. Cell. Commun. Signal. 11:80. 10.1186/1478-811X-11-80 PubMed DOI PMC

Zhou J., Schmid T., Frank R., Brüne B. (2004). PI3K/Akt is required for heat shock proteins to protect hypoxia-inducible factor 1alpha from pVHL-independent degradation. J. Biol. Chem. 279, 13506–13513. 10.1074/jbc.M310164200 PubMed DOI

Ziello J. E., Jovin I. S., Huang Y. (2007). Hypoxia-Inducible Factor (HIF)-1 regulatory pathway and its potential for therapeutic intervention in malignancy and ischemia. Yale J. Biol. Med. 80, 51–60. PubMed PMC

Enlisting the Ixodes scapularis Embryonic ISE6 Cell Line to Investigate the Neuronal Basis of Tick-Pathogen Interactions

Tick galactosyltransferases are involved in α-Gal synthesis and play a role during Anaplasma phagocytophilum infection and Ixodes scapularis tick vector development

Use of Graph Theory to Characterize Human and Arthropod Vector Cell Protein Response to Infection With Anaplasma phagocytophilum

Ixodes scapularis Tick Cells Control Anaplasma phagocytophilum Infection by Increasing the Synthesis of Phosphoenolpyruvate from Tyrosine

Anaplasma phagocytophilum MSP4 and HSP70 Proteins Are Involved in Interactions with Host Cells during Pathogen Infection